The SMARTer Human TCR a/b Profiling Kit provides a powerful new solution for those seeking to perform T-cell receptor (TCR) repertoire analysis using NGS. The kit employs a 5’ RACE-like approach to capture complete V(D)J variable regions of TCR transcripts, starting from human blood RNA samples (10 ng–3 µg) or directly from intact lymphocytes (50–10,000 purified T cells). As the name suggests, the kit can be used to generate data for both TCR-alpha and TCR-beta chain diversity, either in the same experiment or separately. In contrast with TCR profiling methods that involve PCR multiplexing, each round of library amplification utilizes a single primer pair for each TCR subunit. Libraries produced with the kit are indexed and ready for sequencing on Illumina® platforms.

This kit leverages our proven SMART technology (Switching Mechanism at 5’ End of RNA Template) along with LNA technology to amplify TCR mRNA sequences in an unbiased manner with remarkable sensitivity.

Cat. #

Product

Size

Price

License

Quantity

Details

635016

SMARTer® Human TCR a/b Profiling Kit

96 Rxns

$4915.00

The SMARTer Human TCR a/b Profiling Kit enables users to analyze T-cell receptor (TCR) diversity from input RNA samples. This kit is designed to work with a range of RNA input amounts depending on the sample type, and has been shown to generate high-quality libraries from as little as 10 ng to 3 μg of total RNA obtained from peripheral blood leukocytes, or from 50 to 10,000 purified T cells. As the name suggests, the kit can be used to generate data for both alpha- and beta-chain diversity, either in the same experiment or separately.

This kit leverages SMART technology and employs a 5' RACE-like approach to capture complete V(D)J variable regions of TCR transcripts. Included in the kit are primers that incorporate Illumina-specific adaptor sequences during cDNA amplification. The protocol generates indexed libraries that are ready for sequencing on Illumina platforms.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach. Panel A. Reverse transcription and PCR amplification of TCR subunit mRNA sequences. First-strand cDNA synthesis is primed by the TCR dT Primer and performed by an MMLV-derived reverse transcriptase (RT). Upon reaching the 5′ end of each mRNA molecule, the RT adds non-templated nucleotides to the first-strand cDNA. The SMART-Seq v4 Oligonucleotide contains a sequence that is complementary to the non-templated nucleotides added by the RT, and hybridizes to the first-strand cDNA. In the template-switching step, the RT uses the remainder of the SMART-Seq v4 Oligonucleotide as a template for the incorporation of an additional sequence on the end of the first-strand cDNA. Full-length variable regions of TCR cDNA are selectively amplified by PCR using primers that are complementary to the oligonucleotide-templated sequence (SMART Primer 1) and the constant region(s) of TCR-α and/or TCR-β subunits (TCR a/b Human Primer 1). A subsequent round of PCR is performed to further amplify variable regions of TCR-α and/or TCR-β subunits and incorporate adapter sequences, using TCR Primer 2 and TCR a/b Human Primer 2. Included in the primers are adapter and index sequences (read 2 + i7 + P7 and read 1 + i5 + P5, respectively) that are compatible with the Illumina sequencing platform. Following purification, size selection, and quality analysis, the TCR cDNA library is ready for sequencing. Panel B. Semi-nested PCR approach for amplification of TCR-α and/or TCR-β subunits. The primer pairs used for the first round of amplification capture the entire variable region(s) and most of the constant region(s) of TCR-α and/or TCR-β cDNA. The second round of amplification retains the entire variable region(s) of TCR-α and/or TCR-β cDNA, and a smaller portion of the constant region(s). The anticipated size of final TCR library cDNA (inserts + adapters) is ~700–800bp.

635015

SMARTer® Human TCR a/b Profiling Kit

48 Rxns

$3072.00

The SMARTer Human TCR a/b Profiling Kit enables users to analyze T-cell receptor (TCR) diversity from input RNA samples. This kit is designed to work with a range of RNA input amounts depending on the sample type, and has been shown to generate high-quality libraries from as little as 10 ng to 3 μg of total RNA obtained from peripheral blood leukocytes, or from 50 to 10,000 purified T cells. As the name suggests, the kit can be used to generate data for both alpha- and beta-chain diversity, either in the same experiment or separately.

This kit leverages SMART technology and employs a 5' RACE-like approach to capture complete V(D)J variable regions of TCR transcripts. Included in the kit are primers that incorporate Illumina-specific adaptor sequences during cDNA amplification. The protocol generates indexed libraries that are ready for sequencing on Illumina platforms.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach. Panel A. Reverse transcription and PCR amplification of TCR subunit mRNA sequences. First-strand cDNA synthesis is primed by the TCR dT Primer and performed by an MMLV-derived reverse transcriptase (RT). Upon reaching the 5′ end of each mRNA molecule, the RT adds non-templated nucleotides to the first-strand cDNA. The SMART-Seq v4 Oligonucleotide contains a sequence that is complementary to the non-templated nucleotides added by the RT, and hybridizes to the first-strand cDNA. In the template-switching step, the RT uses the remainder of the SMART-Seq v4 Oligonucleotide as a template for the incorporation of an additional sequence on the end of the first-strand cDNA. Full-length variable regions of TCR cDNA are selectively amplified by PCR using primers that are complementary to the oligonucleotide-templated sequence (SMART Primer 1) and the constant region(s) of TCR-α and/or TCR-β subunits (TCR a/b Human Primer 1). A subsequent round of PCR is performed to further amplify variable regions of TCR-α and/or TCR-β subunits and incorporate adapter sequences, using TCR Primer 2 and TCR a/b Human Primer 2. Included in the primers are adapter and index sequences (read 2 + i7 + P7 and read 1 + i5 + P5, respectively) that are compatible with the Illumina sequencing platform. Following purification, size selection, and quality analysis, the TCR cDNA library is ready for sequencing. Panel B. Semi-nested PCR approach for amplification of TCR-α and/or TCR-β subunits. The primer pairs used for the first round of amplification capture the entire variable region(s) and most of the constant region(s) of TCR-α and/or TCR-β cDNA. The second round of amplification retains the entire variable region(s) of TCR-α and/or TCR-β cDNA, and a smaller portion of the constant region(s). The anticipated size of final TCR library cDNA (inserts + adapters) is ~700–800bp.

635014

SMARTer® Human TCR a/b Profiling Kit

12 Rxns

$750.00

The SMARTer Human TCR a/b Profiling Kit enables users to analyze T-cell receptor (TCR) diversity from input RNA samples. This kit is designed to work with a range of RNA input amounts depending on the sample type, and has been shown to generate high-quality libraries from as little as 10 ng to 3 μg of total RNA obtained from peripheral blood leukocytes, or from 50 to 10,000 purified T cells. As the name suggests, the kit can be used to generate data for both alpha- and beta-chain diversity, either in the same experiment or separately.

This kit leverages SMART technology and employs a 5' RACE-like approach to capture complete V(D)J variable regions of TCR transcripts. Included in the kit are primers that incorporate Illumina-specific adaptor sequences during cDNA amplification. The protocol generates indexed libraries that are ready for sequencing on Illumina platforms.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach

Library preparation workflow and PCR strategy for TCR profiling using the SMARTer approach. Panel A. Reverse transcription and PCR amplification of TCR subunit mRNA sequences. First-strand cDNA synthesis is primed by the TCR dT Primer and performed by an MMLV-derived reverse transcriptase (RT). Upon reaching the 5′ end of each mRNA molecule, the RT adds non-templated nucleotides to the first-strand cDNA. The SMART-Seq v4 Oligonucleotide contains a sequence that is complementary to the non-templated nucleotides added by the RT, and hybridizes to the first-strand cDNA. In the template-switching step, the RT uses the remainder of the SMART-Seq v4 Oligonucleotide as a template for the incorporation of an additional sequence on the end of the first-strand cDNA. Full-length variable regions of TCR cDNA are selectively amplified by PCR using primers that are complementary to the oligonucleotide-templated sequence (SMART Primer 1) and the constant region(s) of TCR-α and/or TCR-β subunits (TCR a/b Human Primer 1). A subsequent round of PCR is performed to further amplify variable regions of TCR-α and/or TCR-β subunits and incorporate adapter sequences, using TCR Primer 2 and TCR a/b Human Primer 2. Included in the primers are adapter and index sequences (read 2 + i7 + P7 and read 1 + i5 + P5, respectively) that are compatible with the Illumina sequencing platform. Following purification, size selection, and quality analysis, the TCR cDNA library is ready for sequencing. Panel B. Semi-nested PCR approach for amplification of TCR-α and/or TCR-β subunits. The primer pairs used for the first round of amplification capture the entire variable region(s) and most of the constant region(s) of TCR-α and/or TCR-β cDNA. The second round of amplification retains the entire variable region(s) of TCR-α and/or TCR-β cDNA, and a smaller portion of the constant region(s). The anticipated size of final TCR library cDNA (inserts + adapters) is ~700–800bp.

More Information

Applications

Human TCR repertoire analysis (TCR-alpha and TCR-beta subunits)

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

Takara Bio USA, Inc. (TBUSA, formerly known as Clontech Laboratories, Inc.) provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES (EXCEPT AS SPECIFICALLY NOTED).

Capturem Trypsin provides rapid, efficient, and complete digestion of protein samples, allowing an uninterrupted mass spectometry workflow at room temperature for downstream protein analysis. This product utilizes our novel Capturem technology in a spin column format with membrane-immobilized trypsin. Capturem Trypsin Columns may be used to completely digest protein samples in less than a minute with digestion efficiencies (protein coverage) comparable to or better than those obtained using in-solution trypsin digestion.

Takara Bio USA, Inc. (TBUSA, formerly known as Clontech Laboratories, Inc.) provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES (EXCEPT AS SPECIFICALLY NOTED).